cloning human embryonic stem cells

Posted on by

Image from ScienceNews

Nature reports on the recent success in cloning human embryonic stem cells:

Mitalipov and his group began work on their new study last September, using eggs from young donors recruited through a university advertising campaign. In December, after some false starts, cells from four cloned embryos that Mitalipov had engineered began to grow. “It looks like colonies, it looks like colonies,” he kept thinking. Masahito Tachibana, a fertility specialist from Sendai, Japan, who is finishing a 5-year stint in Mitalipov’s laboratory, nervously sectioned the 1-millimetre-wide clumps of cells and transferred them to new culture plates, where they continued to grow — evidence of success. Mitalipov cancelled his holiday plans. “I was happy to spend Christmas culturing cells,” he says. “My family understood.”

The success came through minor technical tweaks. The researchers used inactivated Sendai virus (known to induce fusion of cells) to unite the egg and body cells, and an electric jolt to activate embryo development. When their first attempts produced six blastocysts but no stable cell lines, they added caffeine, which protects the egg from premature activation.

None of these techniques is new, but the researchers tested them in various combinations in more than 1,000 monkey eggs before moving on to human cells. “They made the right improvements to the protocol,” says Egli. “It’s big news. It’s convincing. I believe it.”

The experiments took only a few months, Mitalipov says. “People say, you did it in monkeys in 2007. Why did it take six years in humans?” Most of the time, he says, was spent navigating US regulations on embryo research.

The researchers carried out a battery of tests to prove that their SCNT cells could form various cell types, including heart cells that are able to contract spontaneously.

Their first cell lines were created using fetal skin cells; others were derived using donor cells from an 8-month-old patient with a rare metabolic disorder called Leigh syndrome, to prove that ESCs could be made from more mature donor cells. The technique does not require prohibitive numbers of eggs: it took 15 from one donor to produce one cell line and 5 from a different donor to make another. “The efficiency was the most impressive thing,” says George Daley, a stem-cell expert at Children’s Hospital Boston in Massachusetts.

Share

fighting tuberculosis with rats

Posted on by

Image from NPR. Abdullah Mchumvu trains African giant pouched rats in Morogoro, Tanzania

Tuberculosis still kills 1.4 million people a year, mostly in the developing world. So it is still beneficial to create new diagnostic techniques, especially when they can be used in rural communities. NPR recently reported on a team of scientists who train African giant pouch rats to sniff out the bacterium in patients’ sputum:

The team trains the critters with a Pavlovian click-and-reward approach. When the rats are just a few weeks old, technicians teach the animals to associate a click sound with a small bite of mashed bananas and a special pellet of food. The next step is to link the scent of TB with the reward.

A trained rat can correctly pick out a TB sample about two-thirds of the time, Beyene says. The rate increases to about 80 percent when two or three animals are put on the task.

The rats aren’t as good as a trained pathologist in the U.S. with a microscope, but they get better results than many clinicians working in rural Africa can achieve, Beyene says. “In an African setting, the sensitivity of the microscopy ranges between 30 to 40 percent,” he explains.

So far APOPO only has around 32 rats in their TB program.

Currently the rats are being used to verify positive test results obtained from microscopic samples.

Share

AIDS vaccine fails

Posted on by

Scanning electron micrograph of HIV-1 budding (in green) from cultured lymphocyte

The past two weeks brought plenty of interesting science news, including the failure of a potential AIDS vaccine. From Popular Science:

The study, called HVTN-505, was begun in 2009, over the years enrolling over 2,500 volunteers. The vaccination process doesn’t actually involve any live or even deactivated HIV; instead, it starts with one that includes genetic material that’s simply modeled after the virus, to prime the immune system. Then comes the real vaccine, involving recombinant DNA (meaning, DNA from various sources) based on adenovirus type 5, a common cold virus that in this case has been disabled so it doesn’t actually cause a cold. Attached to those adenoviruses are artificial versions of HIV antigens. Antigens–the term is short for antibody generator–trigger an immune response, and these artificial antigens were designed to attack the three major HIV subtypes.

This technique had shown some mild success before; in a study in Thailand in 2009, it showed a 31 percent reduction in the HIV infection rate, which sounds good to me, but is apparently not enough to really do more than encourage further research. Unfortunately, that was as much success as this strategy ever saw.

Share

creating oligodendrocytes

Posted on by

Oligodendrocyte

Popular Science covers two papers that appear in Nature Biotechnology on the topic of creating new brain cells. Researchers have developed a method to take skin cells from mice and rats and turn them into oligodendrocytes, which are the type of cells damaged by multiple sclerosis and other disorders.

From Popular Science:

The type of cell that the researchers made is a young, immature version of an oligodendrocyte. Oligodendrocytes normally wrap the nerve fibers of the brain and spinal cord in a protective coating called myelin. With certain diseases, though, people lose that coating or suffer damage to it, which can lead to severe symptoms, such as losing control of the arms and legs.

One major idea researchers have for curing such diseases is adding myelin back by transplanting young, immature oligodendrocytes into the patient. The cells are then supposed to mature and wrap themselves around exposed nerve fibers they find. (Older, more mature oligodendrocytes don’t seem as prone to finding and sheathing exposed nerve fibers.) The idea has worked in lab animals genetically engineered to not have myelin—wohoo!—but there’s a drawback. Until now, researchers generally made oligodendrocytes from stem cells taken from embryos. That’s fine for mice and rats, but it’s difficult to harvest and grow enough embryonic human stem cells for transplants in people.

Share

HIV testing by DVD

Posted on by

A recent paper in Lab on a Chip describes using a DVD player as a diagnostic tool. The researchers convert the optical drive into a laser scanning device that can count the number of CD4+ cells. From Phys Org:

Aman Russom, senior lecturer at the School of Biotechnology at KTH Royal Institute of Technology in Stockholm, says that his research team converted a commercial DVD drive into a laser scanning microscope that can analyse blood and perform cellular imaging with one-micrometre resolution. The breakthrough creates the possibility of an inexpensive and simple-to-use tool that could have far-reaching benefits in health care in the developing world.
“With an ordinary DVD player, we have created a cheap analytical tool for DNA, RNA, proteins and even entire cells,” says Russom. The so-called “Lab-on-DVD” technology makes it possible to complete an HIV test in just a few minutes, he says.
In a proof of concept demonstration, the researchers collected cell-type CD4 + from blood and visualized it using the DVD reader technology. Enumeration of these cells using flow cytometry is now standard in HIV testing, but the practice has been limited in developing countries.
HIV testing currently uses flow cytometry, which requires expensive equipment. If the DVD technique proves reliable, HIV testing can be done much more cheaply.
Share